LOCUS       sequence-211778-        6647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZBTB17 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6647)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6647)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pZBTB17-donor
FEATURES             Location/Qualifiers
     source          1..6647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..49)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(243..262)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     362..384
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(422..440)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        508..612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(613..681,682..1473)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(831..850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1644..2232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2133..2152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2386..2403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2520..2541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2556..2561,2562..2579,2580..2586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2594..2610
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2599..2621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2618..2634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2618..2634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2972..2983
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3433..3453
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3466..3510
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3550..3573
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3601..3603,3604..3606,3607..4320)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3646..3667)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3907..3926)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4358..4931,4932..4934,4935..4943)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4524..4541)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4751..4770
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5025..5828
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5088..5107)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5698..5717
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggtaccgagc tcgaattcac tgccgtcgtt ttacaacgtc gtgactggga aaaccctggc
       61 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
      121 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
      181 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
      241 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
      301 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
      361 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
      421 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
      481 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1141 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1321 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2281 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2341 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2401 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2461 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2521 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2581 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     2641 tacgccaagc ttccttacct gtgtgataag tgtgggcgtg gcttcaaccg ggtagacaac
     2701 ctgcgctccc acgtgaagac cgtgcaccag ggcaaggcag gcatcaagat cctggagccc
     2761 gaggagggca gtgaggtcag cgtggtcact gtggatgaca tggtcacgct ggctaccgag
     2821 gcactggcag cgacagccgt cactcagctc acaggtgcgg gctgtgggca gcagagggaa
     2881 gtcacaggca gcttgctgcc agggtggcga tgatggggca gtgcccgtgc cttcacctta
     2941 ggccctgcga ctctgcccgc agtggtgccg gtgggagctg cagtgacagc cgatgagacg
     3001 gaagtcctga aggccgagat cagcaaagct gtgaagcaag tgcaggaaga aggtgagggg
     3061 ggccacccgg gtgtgaatgt gggcagagcc cggggtcctt cccactctga ctgctgcccc
     3121 acctccacag accccaacac tcacatcctc tacgcctgtg actcctgtgg ggacaagttt
     3181 ctggatgcca acagcctggc tcagcatgtg cgaatccaca cagcccaggc actggtcatg
     3241 ttccagacag acgcggactt ctatcagcag tatgggccag gtggcacgtg gcctgccggg
     3301 caggtgctgc aggctgggga gctggtcttc cgccctcgcg acggggctga gggccagccc
     3361 gcactggcag agacctcccc tacagctcct gaatgtcccc cgcctgccga ggggattcca
     3421 actactgcaa gcgagaatct ttattttcag ggcgccgcca aattcaaaga aaccgctgct
     3481 gctaaattcg aacgccagca catggacagc ggaggtggag gttcgagcgg tccctcgggt
     3541 tcgtcgagcc tggaagttct gttccagggg cccctgtcgt cgagcggtcc ctcgggttcg
     3601 atggtgagca aaggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3661 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3721 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3781 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3841 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3901 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3961 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4021 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     4081 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     4141 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     4201 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4261 cttaaggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     4321 tgatctagag tcgagttaat taagaattcc gccccccccc ctctccctcc ccccccctaa
     4381 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     4441 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     4501 gagcattcct aggggtcttt cccctctcgc caaaggaatg cagggtctgt tgaatgtcgt
     4561 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     4621 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     4681 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     4741 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     4801 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     4861 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4921 acgatgataa tatggccaca acctgttaca ttgcacaaga taaaaatata tcatcacgaa
     4981 cagtaaaact gtctgcttac ataaacagta atacaagggg tgttatggga tcggccattg
     5041 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5101 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5161 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5221 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5281 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5341 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5401 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5461 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5521 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5581 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5641 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5701 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5761 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5821 tcttctgagc tggcggccct tctgactgtt tatttaagga tggatggcac cctggaaccg
     5881 ggaagggtgg cctgttccct agagagaata aattggatta ttttctaatg ctgcctatag
     5941 gtccctgtgg ggtgggtggg gagctggcag ctacaactga tgccacagaa aggctgtacc
     6001 acccaggaac cagccttcct gggaaggtca gcccgtccct cttgctgaat gcgactagaa
     6061 tcagagccag gctggctgct ttgccccaag ttgaggagtg ggcagtacac tcaggctgga
     6121 gagacctcgg caccacgtgc cccatgccct catcccatct tgtttggagg cttctcacct
     6181 gttcatcttt agggcctggc ctcccctcgc cccacagcta tgaccctgct ccccctcggc
     6241 ccaggttggg gagaggttgg gaagttgggg aagggaccag caggggcggc caccaactgc
     6301 ttaccagggt acaggcaacc cagagcgagc ctggctcaca ggccactgct gccaggcctg
     6361 gctcaggggt ccttccagcc agagtggcaa ggtaggccaa ggagagtgcc aggcccaggc
     6421 cactgccctg gctctgatgg gcctaaggga gcagatggaa gccacacttg cttcagggtt
     6481 acctggcccg aggactaagg ccaagccggt gccatcatct tttgggaagg ttcaaaagat
     6541 ggcatctctg ccaccaggct gggggcccag agggacagga aggagcagcc aggggctttg
     6601 atggggaggt agggagctgg tggaacagct ttctcactcc tgtcagc
//
